Genome Engineering with CRISPR

IDT offers 89 modifications that are not listed in our online catalog. A few of the more popular ones are described along with information on how to order them. IDT will consider any modification you have in mind. Please make such inquiries to noncat@idtdna.com.

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) are sequences that serve as an adaptive defense ("immune") systems in bacteria and archaea . In conjunction with CRISPR associated (Cas) proteins, CRISPR sequences are able to recognize and cleave complementary DNA sequences. This natural mechanism has been coopted by scientists for targeted gene editing or removal. This article describes some of the early applications for which this technology is being used.

Exon numbering and location data can differ across various software tools, including with NCBI's gene database. For example, exons within alternatively spliced transcripts are sometimes individually numbered, with no consistent gene-based numbering system across these transcripts for identifying exons. Here we describe how IDT exon location information gives each exon a unique number and how that compares with the NCBI naming system.

Use of the ZEN Quencher as a second, internal quencher in qPCR 5’-nuclease assay probes provides greater overall dye quenching, lowering background, and increasing signal detection. When incorporated into oligonucleotides, it also serves to strengthen duplex formation and block exonuclease digestion, while remaining nontoxic to cells. Thus the ZEN Quencher can be useful in steric blocking antisense oligonucleotide applications.

When performing real-time qPCR, one has to decide whether to use a one-step protocol that combines the RT reaction and PCR in one tube, or a two-step protocol where the RT reaction is performed separately from the PCR.

RNA interference (RNAi) is a conserved pathway found in most eukaryotes where dsRNAs suppress expression of genes with complementary sequences. RNAi has become the experimental tool of choice for studying the effects of gene silencing.

The Clinical and Commercial Manufacturing (CCM) suite is a “facility within a facility” dedicated to the GMP manufacture of oligos for diagnostic use. The CCM suite services customers who need defined specifications or require control over manufacturing processes.

Biotechnology has an inclination towards co-opting nature’s most useful tools. The naturally occurring, extraordinarily strong bond that forms between avidins and biotin has provided a useful tool that has enabled numerous techniques that would otherwise be impossible.

Oligonucleotide synthesis is a complex process that requires more than one hundred sequential chemical reactions to make a single, 25-base sequence. Contemporary synthesis chemistry is robust and modern synthesis platforms are reliable and highly automated. Still, each oligonucleotide synthesized at IDT is evaluated for quality before shipping to ensure that the correct sequence was made. The best method available to assess compound identity in a high throughput environment is mass spectrometry (MS).

Thiol-modified oligonucleotides are used in attachment chemistry reactions to bind an oligo to a target. Targets are commonly gold, but can also include a variety of fluorescent and nonfluorescent moieties.

Cholesterol-TEG is available from IDT for both DNA and RNA oligos on scales from 100 nmole to 10 μmole. Because cholesterol is difficult to manufacture, it requires HPLC purification. To order, select Cholesterol-TEG from the modifications tab on the oligo order page.